DNA methylation patterns of ulcer-healing genes associated with the normal gastric mucosa of gastric cancers.

Recent evidence suggests that gastric mucosal injury induces adaptive changes in DNA methylation. In this study, the methylation status of the key tissue-specific genes in normal gastric mucosa of healthy individuals and cancer patients was evaluated. The methylation-variable sites of 14 genes, including ulcer-healing genes (TFF1, TFF2, CDH1, and PPARG), were chosen from the CpG-island margins or non-island CpGs near the transcription start sites. The healthy individuals as well as the normal gastric mucosa of 23 ulcer, 21 non-invasive cancer, and 53 cancer patients were examined by semiquantitative methylation-specific polymerase chain reaction (PCR) analysis. The ulcer-healing genes were concurrently methylated with other genes depending on the presence or absence of CpG-islands in the normal mucosa of healthy individuals. Both the TFF2 and PPARG genes were frequently undermethylated in ulcer patients. The over- or intermediate-methylated TFF2 and undermethylated PPARG genes was more common in stage-1 cancer patients (71%) than in healthy individuals (10%; odds ratio [OR], 21.9) and non-invasive cancer patients (21%; OR, 8.9). The TFF2-PPARG methylation pattern of cancer patients was stronger in the older-age group (> or =55 yr; OR, 43.6). These results suggest that the combined methylation pattern of ulcer-healing genes serves as a sensitive marker for predicting cancer-prone gastric mucosa.

DNA methylation and expression patterns of key tissue-specific genes in adult stem cells and stomach tissues.

CpG-island margins and non-island-CpG sites round the transcription start sites of CpG-island-positive and -negative genes are methylated to various degrees in a tissue-specific manner. These methylation-variable CpG sites were analyzed to delineate a relationship between the methylation and transcription of the tissue-specific genes. The level of tissue-specific transcription was estimated by counting the number of the total transcripts in the SAGE (serial analysis of gene expression) database. The methylation status of 12 CpG-island margins and 21 non-island CpG sites near the key tissue-specific genes was examined in pluripotent stromal cells obtained from fat and bone marrow samples as well as in lineage-committed cells from marrow bulk, stomach, colon, breast, and thyroid samples. Of the 33 CpG sites examined, 10 non-island-CpG sites, but none of the CpG-island margins were undermethylated concurrent with tissue-specific expression of their nearby genes. The net methylation of the 33 CpG sites and the net amount of non-island-CpG gene transcripts were high in stomach tissues and low in stromal cells. The present findings suggest that the methylation of the non-island-CpG sites is inversely associated with the expression of the nearby genes, and the concert effect of transitional-CpG methylation is linearly associated with the stomach-specific genes lacking CpG-islands.

Chromosomal losses are associated with hypomethylation of the gene-control regions in the stomach with a low number of active genes.

Transitional-CpG methylation between unmethylated promoters and nearby methylated retroelements plays a role in the establishment of tissue-specific transcription. This study examined whether chromosomal losses reducing the active genes in cancers can change transitional-CpG methylation and the transcription activity in a cancer-type-dependent manner. The transitional-CpG sites at the CpG-island margins of nine genes and the non-island-CpG sites round the transcription start sites of six genes lacking CpG islands were examined by methylation-specific polymerase chain reaction (PCR) analysis. The number of active genes in normal and cancerous tissues of the stomach, colon, breast, and nasopharynx were analyzed using the public data in silico. The CpG-island margins and non-island CpG sites tended to be hypermethylated and hypomethylated in all cancer types, respectively. The CpG-island margins were hypermethylated and a low number of genes were active in the normal stomach compared with other normal tissues. In gastric cancers, the CpG-island margins and non-island-CpG sites were hypomethylated in association with high-level chromosomal losses, and the number of active genes increased. Colon, breast, and nasopharyngeal cancers showed no significant association between the chromosomal losses and methylation changes. These findings suggest that chromosomal losses in gastric cancers are associated with the hypomethylation of the gene-control regions and the increased number of active genes.

The 5'-end transitional CpGs between the CpG islands and retroelements are hypomethylated in association with loss of heterozygosity in gastric cancers.

BACKGROUND: A loss of heterozygosity (LOH) represents a unilateral chromosomal loss that reduces the dose of highly repetitive Alu, L1, and LTR retroelements. The aim of this study was to determine if the LOH events can affect the spread of retroelement methylation in the 5'-end transitional area between the CpG islands and their nearest retroelements. METHODS: The 5'-transitional area of all human genes (22,297) was measured according to the nearest retroelements to the transcription start sites. For 50 gastric cancer specimens, the level of LOH events on eight cancer-associated chromosomes was estimated using the microsatellite markers, and the 5'-transitional CpGs of 20 selected genes were examined by methylation analysis using the bisulfite-modified DNA. RESULTS: The extent of the transitional area was significantly shorter with the nearest Alu elements than with the nearest L1 and LTR elements, as well as in the extragenic regions containing a higher density of retroelements than in the intragenic regions. The CpG islands neighbouring a high density of Alu elements were consistently hypomethylated in both normal and tumor tissues. The 5'-transitional methylated CpG sites bordered by a low density of Alu elements or the L1 and LTR elements were hypomethylated more frequently in the high-level LOH cases than in the low-level LOH cases. CONCLUSION: The 5'-transitional methylated CpG sites not completely protected by the Alu elements were hypomethylated in association with LOH events in gastric cancers. This suggests that an irreversible unbalanced decrease in the genomic dose reduces the spread of L1 methylation in the 5'-end regions of genes.

Determinants of respiratory symptom development in patients with chronic airflow obstruction.

BACKGROUND: This study was undertaken to identify the determinants of respiratory symptom development in patients with chronic airflow obstruction (CAO). METHODS: Categories of symptomatic and asymptomatic CAO were defined using questionnaire responses and spirometric results. We analyzed data obtained as part of the second South Korean National Health and Nutrition Examination Survey (Korean NHANES II). RESULTS: Among 187 patients with CAO, 69 had no respiratory symptoms. CAO patients with symptoms were significantly older than those without symptoms (P=0.026), and hypertension was more common among symptomatic CAO patients than among asymptomatic CAO patients (P=0.005). According to questionnaire responses, symptomatic CAO patients had more difficulty in walking or lifting (P<0.001), required more help with personal care (P=0.01), and had poorer general health than asymptomatic CAO patients (P=0.008). Symptomatic CAO patients had higher fasting blood glucose levels than asymptomatic CAO patients (P=0.028). Symptomatic CAO patients had significantly lower forced expiratory volume in 1s (FEV1) (P=0.001), forced vital capacity (FVC) (P=0.008), and a ratio of FEV1/FVC than asymptomatic CAO patients (P<0.001). Statistically significant predictors of symptom development were as follows: age (odds ratio (OR) 1.04, P=0.028), hypertension (OR 4.41, P=0.008), fasting blood glucose (OR 1.02, P=0.034), FEV1 (OR 0.07, P=0.002), FVC (OR 0.08, P=0.009), FEV1/FVC (OR 0.00, P=0.001). Multiple logistic regression analyses revealed two independent factors associated with symptom development: FEV1/FVC (OR 0.001, P=0.002) and hypertension (OR 5.95, P=0.005). CONCLUSIONS: In CAO, respiratory symptom development is significantly associated with low FEV1/FVC and the presence of hypertension.

A case of gastroenteritis associated with gastric trichuriasis.

A rare human case of gastroenteritis and eosinophilic ascites associated with gastric trichuriasis is described. The patient was a 32-yr-old woman who was working in a farm near Pohang, Korea. She complained of abdominal pain, diarrhea, and vomiting. Endoscopic examination found focal linear hyperemia on the mucosa of the stomach antrum, and endoscopic biopsy confirmed eosinophilic inflammation of the mucosa and submucosa of the stomach, terminal ileum, and cecum. The biopsy specimen of the stomach included a female Trichuris trichiura which was covered by many inflammatory cells on its surface. Ascites and intestinal wall thickening was found by CT scan, and Douglas pouch centesis aspirated bloody ascites which included many eosinophils. She was medicated with prednisolone and albendazole and cured. She is the first case of eosinophilic inflammation of the gastrointestinal tract and ascites associated with trichuriasis in the stomach.

A large malignant insulinoma: case report with endosonographic, immunohistochemical and ultrastructural features.

Malignant insulinoma in the beta cells of the pancreatic islet is rare and usually presented as hypoglycemia. We report a case of large malignant insulinoma in a 53-year-old Korean woman. A presumptive clinical diagnosis was made before surgery, based on the high plasma insulin-to-glucose ratio and a large solitary heterogeneous pancreatic mass by abdominal computed tomography and endosonography. The tumor measured 5.8 x 4.7 x 4.5 cm in dimension and showed capsular invasions and metastases in two of four peripancreatic lymph nodes. The tumor cells were strongly immunoreactive to insulin and had a high Ki-67 labeling index (13%) and atypical membranous electron-dense granules, ranging from 120 to 400 nm in diameter, in the cytoplasm on electron microscopy. The patient was treated by distal pancreatectomy with splenectomy and rapidly recovered without neurohypoglycemic symptoms. This case showed not only lymph node metastases, the most reliable parameter for malignancy in pancreatic endocrine tumors, but also other valid diagnostic clues, such as high Ki-67 labeling index, heterogeneous enodosonographic findings, capsular invasions with large tumor and pure atypical secretory granules.